Professional Documents
Culture Documents
Rules 2019 Chapter 1
Rules 2019 Chapter 1
https://doi.org/10.15258/istarules.2019.01
The electronic version of the International Rules for Seed Testing includes the English,
French, German and Spanish versions. If there are any questions on interpretation of the
ISTA Rules, the English version is the definitive version.
Published by
The International Seed Testing Association (ISTA)
Zürichstr. 50, CH-8303 Bassersdorf, Switzerland
All rights reserved. No part of this publication may be reproduced, stored in any retrieval
system or transmitted in any form or by any means, electronic, mechanical, photocopying,
recording or otherwise, without prior permission in writing from ISTA.
Contents
Preface to the 2019 Edition of the ISTA Rules .............. v 1.5.2.20 Seed mixtures .......................................... 1-11
1.5.2.20.1 Purity and component analysis .......... 1-11
Chapter 1: ISTA Certificates ....................................... 1-1 1.5.2.20.2 Determination of other seeds by
1.1 Object ....................................................................... 1-1 number ........................................................... 1-11
1.2 Definitions ................................................................ 1-1 1.5.2.20.3 Germination, seed viability, seed
1.2.1 Orange International Seed Lot Certificate ......... 1-1 vigour and other tests using replicates of 100
1.2.2 Blue International Seed Sample Certificate ....... 1-1 seeds ................................................................ 1-11
1.2.3 Original certificate .............................................. 1-1 1.5.2.20.4 Thousand-seed weight ....................... 1-11
1.2.4 Duplicate certificate ........................................... 1-1 1.5.2.21 Genetically modified organisms .............. 1-12
1.2.5 Provisional certificate ......................................... 1-1 1.5.2.21.1 Qualitative test results ....................... 1-12
1.2.6 Accredited laboratory ......................................... 1-1 1.5.2.21.2 Quantitative results obtained by
1.3 Conditions for issuance of ISTA Certificates ........... 1-2 multiple qualitative tests of individuals or
1.4 Completing ISTA Certificates ................................... 1-3 groups of seeds or seedlings .......................... 1-12
1.4.1 General ............................................................... 1-3 1.5.2.21.3 Quantitative measurements of GMO
1.4.2 Orange International Seed Lot Certificate ......... 1-3 in bulk samples .............................................. 1-12
1.4.3 Blue International Seed Sample Certificate ....... 1-3 1.5.2.22 Reporting of results of tests not covered
1.4.4 Duplicate certificate ........................................... 1-4 by the Rules ........................................................ 1-12
1.4.5 Provisional certificate ......................................... 1-4 1.5.3 Reporting of uncertainty of measurement on
1.5 Reporting results ....................................................... 1-4 ISTA Certificates .................................................... 1-12
1.5.1 Sampling and testing .......................................... 1-4 1.5.4 Statement referring to compliance with
1.5.2 Certificates ......................................................... 1-4 legislative requirements ......................................... 1-13
1.5.2.1 Sampling: heterogeneity testing for seed 1.6 Validity of ISTA Certificates ................................... 1-13
lots in multiple containers .................................... 1-4 1.7 Disputed results ...................................................... 1-13
1.5.2.1.1 The H value heterogeneity test .............. 1-4
1.5.2.1.2 The R value heterogeneity test .............. 1-4
1.5.2.2 Purity ........................................................... 1-4
1.5.2.3 Purity tests on coated seeds ......................... 1-5
1.5.2.4 Determination of other seeds by number ..... 1-6
1.5.2.5 Determination of other seeds by number
on coated seeds ..................................................... 1-6
1.5.2.6 Germination ................................................. 1-6
1.5.2.7 Germination of coated seeds ........................ 1-7
1.5.2.8 Tetrazolium test ........................................... 1-7
1.5.2.9 Tetrazolium test on coated seeds ................. 1-7
1.5.2.10 Seed health test .......................................... 1-8
1.5.2.11 Species and variety testing ......................... 1-8
1.5.2.11.1 Results of examination of individual
seeds or seedlings .............................................. 1-8
1.5.2.11.2 Results of a field plot examination ...... 1-9
1.5.2.12 Moisture content ........................................ 1-9
1.5.2.13 Thousand-seed weight ............................... 1-9
1.5.2.14 Excised embryo ......................................... 1-9
1.5.2.15 Weighed replicates ..................................... 1-9
1.5.2.16 X-ray test ................................................... 1-9
1.5.2.17 Seed vigour test ........................................ 1-9
1.5.2.17.1 Conductivity test ................................. 1-9
1.5.2.17.2 Accelerated ageing test ...................... 1-10
1.5.2.17.3 Controlled deterioration test .............. 1-10
1.5.2.17.4 Radicle emergence test ...................... 1-10
1.5.2.17.5 Tetrazolium vigour test ...................... 1-10
Contents
Since 2014, the International Rules for Seed Testing (ISTA Details of changes
Rules) are primarily available in electronic form only. The
ISTA Rules can be downloaded as a complete PDF file or The 2019 changes are editorial corrections or Rules
as individual chapters from: changes adopted at the Ordinary General Meeting held at
http://www.ingentaconnect.com/content/ista/rules Sapporo, Japan, in June 2018. Edits were made in Adobe
If required, users of the ISTA Rules can print their own InDesign by Vanessa Sutcliffe of HeartWood Editorial
copies. For further information on the ISTA Rules, see: (www.heartwoodeditorial.co.uk).
http://www.seedtest.org/rules The changes in the text content from the previous
The electronic version includes the English, French, edition of the ISTA Rules are listed below. They can be
German and Spanish versions of the ISTA Rules. If there displayed as yellow highlighted text as a ‘layer’ within
are any questions on interpretation of the ISTA Rules, the the electronic copy with comments on what has changed.
English version is the definitive version. For the previous history of amendments to the ISTA
Rules, see the Prefaces for 2003 to 2018 on the ISTA
website.
Seed health testing methods
Ernest Allen, ISTA Rules Committee Chair
Previously, the seed health testing methods were published
as a separate Annexe to Chapter 7 of the ISTA Rules. They Susan Alvarez, ISTA Rules Committee Vice-Chair
are now available as separate method sheets from the
ISTA web site at: ISTA Secretariat
http://www.seedtest.org/seedhealthmethods
Chapter 3 Chapter 8
3.7: Consequential changes in clarification of reporting 8.3.3: Sentence added (‘DNA can be extracted... equiva-
on ISTA Certificates. Points removed (‘The percentage lent to one seedling’).
by weight... are listed first.’ and ‘The kinds of inert mat- 8.9.4.3.2, 8.9.6.6.1.2, 8.9.6.6.2.2: Sentence added (‘Note
ter... (to one decimal place).’). that as an alternative... prior to pouring the gel.’) for con-
Table 3B Part 1: New PSD numbers included (for sistency in gel polymerisation method.
Eustoma and Felicia).
Chapter 10
Chapter 4 Change of title from ‘Weight determination’ to ‘Thou-
4.5.3, 4.5.3.1, 4.5.3.2, 4.5.3.3, 4.5.3.4, 4.7: Addition of sand-seed weight (TSW) determination’. Revision to
other species of parasitic weeds with dust-like seed under whole chapter and addition of Table 10A.
‘Determination of other seeds by number’.
4.5.3.1: Sentence added (‘This test is not... pelleted Chapter 11
seed.’). 11.1.1: Cross reference updated under ‘Seed treatment’
to ‘2.2.12’.
Chapter 5 11.2.5.4.1: Duplication of lot size definition deemed un-
5.2.7.2: Addition of criterion on evaluation of the root necessary so only the reference to Chapter 2 is kept.
system of Spinacia oleracea seedlings, supported by a Table 11A: Consequential change in table due to revision
validation study. of Chapter 10.
The results reported on an Orange International Seed An accredited laboratory is an ISTA-accredited member
Lot Certificate refer strictly to the lot as a whole at the laboratory authorised by the ISTA Executive Committee
time of sampling. under Article 4(i) of the Articles of ISTA to sample and
test seeds and to issue ISTA Certificates.
1.3 Conditions for issuance of ISTA e. To report results of tests which are in the ISTA Rules,
Certificates the laboratory must be accredited for these tests, either
directly or through subcontracting to another labora-
ISTA Certificates must be issued only on forms obtained tory accredited for these tests.
from the ISTA Secretariat and approved by the ISTA Ex- f. The assessment of any attribute reported on a certifi-
ecutive Committee. There are two kinds of certificates: cate must be calculated from tests carried out on one
Orange International Seed Lot Certificates and Blue Inter- submitted sample.
national Seed Sample Certificates, as defined in 1.2. g. In the case of Orange International Seed Lot
On request of the applicant, duplicate and provisional Certificates:
certificates as defined in 1.2 may be issued. – the seed lot must comply with the requirements
A duplicate certificate may be issued for an original prescribed in 2.5.4;
certificate. More than one duplicate certificate may be – the submitted sample must be drawn and dealt with
issued. in accordance with 2.5.4.
A provisional certificate may be issued for any ISTA h. For an Orange International Seed Lot Certificate,
test result(s) that are later combined onto an original each container in the lot must be marked, labelled and
certificate. More than one provisional certificate may be sealed in accordance with 2.5.4.3.
issued. i. For an Orange International Seed Lot Certificate to be
If an applicant cancels testing, an ISTA Certificate issued for a sublot, the sublot must represent a mini-
does not need to be issued. mum size of 20 % of the weight of the original seed
lot. A maximum of five Orange International Seed Lot
An ISTA Certificate may be issued only by the seed test- Certificates may be issued for sublots of any one origi-
ing laboratory which either carried out all the tests to be nal seed lot.
reported, or subcontracted sampling and/or some of the j. For an Orange International Seed Lot Certificate, the
tests to be reported (see 1.4.2 and 1.4.3), and under the submitted sample must be tested by an accredited
conditions listed below: laboratory. The issuing laboratory must ensure that
a. The issuing laboratory must be currently authorised to sampling, sealing, identification, testing and issuance
do so by the Executive Committee. of the certificate is in accordance with the ISTA Rules,
b. The seed tested must be of a species listed in Table 2A although subcontracting of sampling and/or testing to
(Lot and sample weights) of the ISTA Rules. Where in another accredited laboratory is permissible. The labo-
other tables, such as Table 5A and Table 6A, methods ratory which carries out sampling must provide all the
are prescribed for a group of species, only those spe- information that is necessary to complete the Orange
cies specifically listed in Table 2A may be considered International Seed Lot Certificate.
to be covered. The seed lot identification (‘Marks of the lot’; see
Consequently, no certificates may be issued for species 2.2.10) may take the form of a sequential series of
not listed in Table 2A of the current ISTA Rules, ex- characters or a single reference character. Each con-
cept in the case of seed mixtures, where for the species tainer within the lot or sublot must be identified in such
tested it is shown as ‘Seed mixture’. a way that the containers can be readily recognised
c. The tests must be carried out in accordance with the by the information provided on the certificate issued.
ISTA Rules. However, additionally and on request, Each container of a sublot must be marked with the
results of tests not covered by these Rules may be re- identification of the original seed lot. A sublot-specific
ported on an ISTA Certificate (see 1.5.2.22). identification is not necessary.
Results of analyses not covered by the current ISTA When the seed lot is located in a different country to
Rules may be included on a certificate only if results the sampling laboratory, the country where the seed lot
Chapter 1: ISTA Certificates
of at least one test covered by the ISTA Rules are also has been sampled must be reported either under ‘Sam-
being reported. pling by’ or under ‘Additional observations’.
d. For the result of a determination of moisture content to
be reported on an ISTA Certificate, the sample must be
submitted in an intact, moisture-proof container from
which as much air as possible has been excluded (see
9.2.5.1).
1.4 Completing ISTA Certificates The completed Orange International Seed Lot Certificate
must show the following information:
1.4.1 General a. name, address, ISTA member code and stamp (seal) of
issuing laboratory;
ISTA Certificates must be completed using a typewriter b. name and ISTA member code of laboratory responsi-
or machine-printer and can be completed in any language. ble for sampling;
No certificate may be issued that shows signs of amend- c. seed lot identification (i.e. marks of lot);
ment, alteration or erasure. d. Under ‘Seal of lot’: the method of sealing (e.g. stitch-
ing, metal seal) and/or the authority (e.g. ISTA labora-
A completed certificate must show the following tory, Ministry).
information: e. either the number of containers for which the certifi-
a. The name and address of the issuing laboratory; the cate is issued; or ‘N/A’ for ‘not applicable’;
laboratory must be on the ISTA list of accredited mem- f. date of sampling;
ber laboratories. g. date that the sample was received by the testing
b. Dates, written in the ISO 8601 format: year in full – laboratory;
month – day, with two figures for both month and day h. date test was concluded;
(e.g. 2007-07-25). i. place, country and date of issue of the certificate;
c. The scientific name of the species tested, as listed in j. test or sample number of the testing laboratory;
the current ISTA Rules and (in most cases) also the k. analysis results;
ISTA List of Stabilised Plant Names. Where it is im- l. In the case of certificates for sublots, under ‘Other de-
possible to determine the species with certainty on the terminations’: ‘The results reported represent the sam-
basis of seed characters, only the genus name must be ple drawn from the original seed lot of … kg’;
stated (example: Malus sp.). In the case of seed mix- m. country where the seed lot was sampled, when the seed
tures, for the species tested ‘Seed mixture’ must be lot is located in a different country to the sampling lab-
entered. oratory, reported under either ‘Sampling by’ or ‘Ad-
d. The name and address of the applicant. Other informa- ditional observations’;
tion stated by the applicant, such as country of origin, n. the signature of the Head of the issuing laboratory or
species, cultivar, weight of lot or sublot, certification their assignee which confirms the statement on the
category and applicant’s lot reference must be entered back of the certificate as true;
as stated by the applicant. o. under the signature it must state at least, the job posi-
Note: at the request of the applicant the name and ad- tion of the person signing or ‘Authorised signatory’.
dress of the applicant may be omitted.
e. The signature of the Head of the issuing laboratory or
their assignee. It may be either a physical or an elec- 1.4.3 Blue International Seed Sample
tronic signature, the use of which is authorised by the Certificate
Head of the issuing laboratory.
f. Under ‘Status of certificate’, the word ‘ORIGINAL’, The Blue International Seed Sample Certificate refers
‘PROVISIONAL’ or ‘DUPLICATE’, as appropriate. only to the sample submitted for testing.
It is stated on the back of the Blue International Seed
Sample Certificate:
1.4.2 Orange International Seed Lot ‘I certify that testing has been carried out in accord-
Certificate ance with the International Rules for Seed Testing of the
ISTA and that the tests have been made at a laboratory ac-
Chapter 1: ISTA Certificates
It is stated on the back of the Orange International Seed credited by the International Seed Testing Association to
Lot Certificate: issue International Seed Analysis Certificates.’
‘I certify that sampling, sealing and testing have been
carried out in accordance with the International Rules The completed certificate must show the following
for Seed Testing of the ISTA and that the tests have been information:
made at a laboratory accredited by the International Seed a. name, address, ISTA member code and stamp (seal) of
Testing Association to issue International Seed Analysis issuing laboratory;
Certificates.’ b. date that the sample was received by the testing
laboratory;
c. date test was concluded;
d. place, country and date of issue of the certificate;
e. test or sample number of the testing laboratory;
A provisional ISTA Certificate may be issued on request Note: the H value must not be calculated or reported if X
of the applicant. There is no limit to the number of copies is outside the following limits:
of an ISTA certificate marked ‘Provisional’. – purity components: above 99.8 % or below 0.2 %;
If the sample is being tested for more than one test, but – germination: above 99.0 % or below 1.0 %;
at least one test has been concluded, the ‘date that the test – number of specified seeds: below two per sample.
was concluded’ is entered.
The results of tests may be reported on one or more ISTA The results of a purity test must be reported in the spaces
Certificates, separately or combined. provided as follows:
Test results must be reported in accordance with the – The scientific name of the species of pure seed, in
rules for calculating, expressing and reporting results accordance with Table 2A (e.g. Triticum aestivum).
Chapter 1: ISTA Certificates
in the appropriate chapter of the ISTA Rules. If there is Where it is impossible to determine the species with
a space on the certificate for a certain test which is not certainty on the basis of seed characteristics, reporting
made, or where information is not provided by the appli- must be done to the most precise taxon possible.
cant: ‘N’ for ‘not tested’ or ‘NA’ for ‘not applicable’, must – The percentage by weight of pure seed, inert matter
be placed in the space. and other seeds, given to one decimal place. The per-
‘N’ must not be used for components of a test such as centage of all components must total 100 %. Compo-
hard or fresh seeds. For example, even if hard seeds do not nents amounting to less than 0.05 % must be reported
usually occur in the species they must be looked for and if as ‘Trace’ or ‘TR’ (for ‘Trace’). If no inert matter or
not found, then reported as ‘0’. other seeds are found, this must be reported as ‘0.0’.
– The kind of inert matter.
– The scientific name of every species of other seeds – Multiple seed units must be reported as ‘% MSU’.
found, in accordance, where applicable, with the cur- – Seeds with appendages attached must be reported as
rent ISTA List of Stabilised Plant Names, available at ‘% seeds with appendages attached’.
www.seedtest.org/stablist (e.g. Elytrigia repens). – The percentage by weight of broken pure seed.
– When the weight of the working sample tested for pu-
rity equals or is no more than 10 % higher than the The percentages may be reported to more than one deci-
weight specified in Table 2A, column 4 (Purity analy- mal place if requested.
sis), no statement regarding the weight of the working
sample is required on the ISTA Certificate.
– When the weight of the working sample tested for pu- 1.5.2.3 Purity tests on coated seeds
rity deviates from that specified in Table 2A, column
4, the actual weight of the working sample weighed The result of a purity test on coated seeds must be reported
according to 3.5.1 must be reported on the ISTA Cer- as follows:
tificate using one of the following, as applicable: – Following the species name, the words ‘seed pellets’,
a. When testing a weight that exceeds by 10 % the ‘encrusted seeds’, ‘seed granules’, ‘seed tapes’ or
weight specified in Table 2A, column 4, report un- ‘seed mats’, as applicable, must be clearly entered.
der other determinations as: – The results must be reported to one decimal place, and
‘Purity: ... g’ the percentage of all components must total 100 %.
b. When testing a weight estimated to contain 2500 Components amounting to less than 0.05 % must be
seed units, report under other determinations as: reported as ‘Trace’ or ‘TR’ (for ‘Trace’). If no inert
‘Purity: …g (approx. 2500 seeds)’ matter or other seeds are found, this must be reported
c. When the submitted sample received for purity as ‘0.0’.
testing weighs less than the weight in Table 2A, – In the case of pelleted seeds only, the percentages of
column 4, report under other determinations and pure pelleted seeds, inert matter and unpelleted seeds
use the current statement, according to 2.5.4.5: must be reported in the spaces provided for ‘Pure
‘The submitted sample weighed only … g and is seeds’, ‘Inert matter’, and ‘Other seeds’, respectively.
not in accordance with the International Rules for – The name and number of the seeds of each species
Seed Testing.’ found in the examination of the 100 seeds removed
– The percentage of winged seed (as defined in Pure from the pellets or tapes must be reported under ‘Other
Seed Definitions 47 and 51), if winged seeds are found. determinations’.
Upon request, the following information can be reported Upon request, the following information may be reported
to one decimal place in the spaces provided for the % pu- under ‘Other determinations’ as follows:
rity results: – Purity test on depelleted seeds. The component parts
– The percentage by weight of a specified species, en- (pure seed, other seeds and inert matter) may be re-
tered immediately after the name of the species to the ported as percentages of their total weight, ignoring
nearest 0.1 %. Species for which the percentage by the pelleting material. The percentage of pelleting ma-
weight has been requested are listed first. terial must be reported separately only on request. The
– The percentage by weight of any kind of inert matter. result of this test is to be reported: ‘weight of … mate-
rial excluded’.
Upon request, the percentages can be reported to more than – Purity of seeds removed from tapes. The component
one decimal place but only under ‘Other determinations’. parts (pure seed, other seeds, and inert matter) may be
reported as percentages of their total weight, ignoring
Chapter 1: ISTA Certificates
Upon request, the following information must be reported the tape material. The result of this test is to be re-
under ‘Other determinations’ as follows: ported: ‘weight of … material excluded’.
– Other seeds may be divided into ‘other crop seeds’
and ‘weed seeds’. In this case, the words ‘Other crop
seeds’ must be entered, followed by the percentage
by weight of other crop seeds and the name(s) of the
species found. This procedure must also be used for
‘Weed seeds’.
The result of a determination of other seeds by number The result of a determination of other seeds by number on
must be reported under ‘Other determinations’ as follows: coated seeds must be reported as follows:
– The actual weight of seed examined to the minimum – Following the species name, the words ‘seed pellets’,
number of decimal places indicated in Table 4.1. ‘encrusted seeds’, ‘seed granules’, ‘seed tapes’ or
– The scientific name and number of seeds of each spe- ‘seed mats’, as applicable, must be clearly entered.
cies sought and found in this weight. If no other seeds – Under ‘Other determinations’, the actual weight (or
are found, this must be indicated on the certificate. length of tape, or area of mat) and approximate num-
– Where it is impossible to determine with certainty on ber of pelleted seeds examined must be entered, to-
the basis of seed characteristics, reporting must be gether with the scientific name and number of seeds of
done to the most precise taxon possible. each species sought and found in this weight, length or
– If the full weight prescribed in Table 2A was examined area.
for all other species present, then the words ‘Complete
test’ must be entered, alongside the weight of seed Upon request, the result may in addition be expressed in
examined. some other way, such as number of seeds per kilogram,
– If the examination was for only a limited range of other per metre or per square metre.
species, then the words ‘Limited test’ must be entered.
– If the weight examined for all other species was less
than the prescribed weight, then the words ‘Reduced 1.5.2.6 Germination
test’ must be entered.
– If the weight examined was less than the weight pre- The result of a germination test must be reported in the
scribed in Table 2A, and only a limited range of other spaces provided as follows:
species was examined, then the words ‘Reduced-limit- – The actual duration of the test (in days, excluding the
ed test’ must be entered. period of special treatment or method used for promot-
– If a sample of at least 25 000 seeds was examined, ing germination).
and this sample was below the weight prescribed in – The percentages, calculated to the nearest whole num-
Table 2A, then the weight of seed examined and the ber (5.8.2), of normal seedlings, hard seeds, fresh
statement ‘Test based on at least 25 000 seeds’ must be seeds, abnormal seedlings and dead seeds. If the result
entered. for any of these categories is found to be zero, it must
be reported as ‘0’.
Upon request, the results may in addition be expressed in – If an applicant requests that the test be terminated
some other way, such as ‘weight of seeds found’ or ‘num- when the sample reaches a predetermined germination
ber of seeds per kilogram’. percentage, before the final count, then only the per-
Upon request, the presence of one or more of the fol- centage of normal seedlings is reported. The results of
lowing genera: Aeginetia, Alectra, Orobanche and Striga, the other categories (abnormal seedlings, hard seeds,
can be reported only on a Blue International Seed Sample fresh seeds and dead seeds) must be reported as ‘N’,
Certificate (see 1.2.2) and must be reported as: ‘Test for because they have not been determined.
presence of ... species – … seeds of ... species were found
in … g of seed examined.’ The following additional information must be reported
If no seeds were found it can be reported as: ‘No seeds under ‘Other determinations’:
of ... species were found in … g of seed examined.’ – the number of seeds tested, if less than 400 seeds;
Chapter 1: ISTA Certificates
The sample weight examined must be reported accord- – the germination method using the abbreviations used in
ing to the minimum number of decimals indicated in Table Table 5A, including at least substrate and temperature;
4.1. – any special treatment or method used for promoting
germination (5.6.3);
– the duration in days of any special treatment or method
used for promoting germination, except in the case of
prestorage;
– the germination percentage obtained within the pre- The following additional information must also be report-
scribed time, if the germination period was extended ed under ‘Other determinations’:
beyond the period indicated in Table 5A. The state- – The method used for the germination test.
ment must be entered as follows: ‘After the prescribed – For seed tapes or mats: the number of normal seed-
period of … days, there were … % normal seedlings’; lings per metre of tape or square metre of mat.
– the method for evaluating fresh seeds (dissection, tetra-
zolium or excised embryo – see paragraph 5.6.5.3.) Seedlings that are obviously not of the species stated by
when 5 % or more of fresh seeds are believed to be the applicant, even if otherwise normal, must not be in-
present; cluded in the germination result, but their number must be
– If an applicant requests that the germination test be ter- reported separately.
minated when the sample reaches a predetermined ger-
mination percentage, the following statement: ‘Upon
request of the applicant, the germination test was ter- 1.5.2.8 Tetrazolium test
minated after … days. The prescribed test period is …
days.’ The result of a tetrazolium test must be reported under
‘Other determinations’ as follows:
When double tests are prescribed in Table 5A Part 2, the – The statement ‘Tetrazolium test: …% of seeds were
result of the first test, with treatment for breaking dor- viable’ must be entered.
mancy, is reported in the appropriate space on the ISTA – In cases where the testing procedure deviates from that
Certificate, and the result of the second test, without treat- prescribed in Table 6A, any deviating procedure must
ment for breaking dormancy, is reported under ‘Other also be reported.
determinations’. The only variations permitted from procedures given
in Table 6A are for premoistening time, tetrazolium
Upon request, the following information may be reported concentration, staining temperature or staining time.
as follows: Precise prescriptions about the limitation of the vari-
– the result of parallel tests or any additional test; ations are given in 6.5.
– the viability of ungerminated seeds and the method – If individual seeds are tested at the end of the germi-
used to determine it; nation test, the result must be reported in accordance
– the categories of ungerminated seeds (as listed in with 1.5.2.6 and 5.9.
5.6.5.3) and the method used to determine them;
– in the case of multigerm seed units: the number of nor- In addition, in the case of species of Fabaceae, one of the
mal seedlings produced by 100 units, the number of following, and only one, must be reported:
units which have produced one, two or more than two either (in cases where the viability percentage of the hard
normal seedlings, or the proportion of units producing seed is not determined) ‘Tetrazolium test: ...% of seeds
one, two or more than two normal seedlings. The pro- were viable, ...% of hard seeds found in the test.’
portion is expressed as a percentage of the total num- or (in cases where the viability percentage of the hard
ber of units which have produced at least one normal seed is determined) ‘Tetrazolium test: ...% of seeds
seedling. were viable, ...% of hard seeds included in the percent-
age of viable seed’
1.5.2.7 Germination of coated seeds At the discretion of the seed testing laboratory, further in-
formation may be reported, e.g. percentage of seeds that
The result of a germination test on coated seeds must be were empty, with larvae, broken or decayed.
Chapter 1: ISTA Certificates
reported as follows:
– Following the species name, the words ‘seed pellets’,
‘encrusted seeds’, ‘seed granules’, ‘seed tapes’ or 1.5.2.9 Tetrazolium test on coated seeds
‘seed mats’, as applicable, must be clearly entered in
the space provided. The result of a tetrazolium test on coated seeds must be
– The percentage of pellets or seed in tapes with nor- reported as follows:
mal seedlings, with abnormal seedlings and without – Following the species name, the words ‘seed pellets’,
seedlings. ‘encrusted seeds’, ‘seed granules’, ‘seed tapes’ or
– The duration of the test. ‘seed mats’, as applicable, must be clearly entered.
The following additional information must be reported 1.5.2.11 Species and variety testing
under ‘Other determinations’:
– The statement ‘Number of seeds (of the species stated The results of species and variety testing must be reported
by the applicant) included in 100 seed pellets’ (or ‘en- under ‘Other determinations’, and in addition the follow-
crusted seeds’, or ‘seed granules’); ing information must be given:
– or the statement ‘Number of seeds (of the species a. the request of the applicant;
stated by the applicant) included in one metre of seed b. the trait(s) and the method(s) used;
tape’; c. the kind of preparation of the working sample (e.g. the
– or the statement ‘Number of seeds (of the species stat- whole working sample excluding the inert matter or
ed by the applicant) included in one seed mat or in one only the pure seed fraction, washing);
square metre of seed mat’. d. whether an authentic standard sample or a standard
– The statement ‘Tetrazolium test: …% were viable’ reference was used; if a standard reference was used,
must be entered. its origin must be indicated;
– In cases where the testing procedure deviates from that e. the number of seeds, seedlings or plants examined.
prescribed in Table 6A, any deviating procedure must When it is difficult to determine the total number of
also be reported. The only areas where variations from plants examined in field plots, the mass of seed sown
procedures given in Table 6A are permitted are for must be reported.
premoistening time, tetrazolium concentration, stain-
ing temperature and staining time. For precise guid-
ance about the limitation of the variations permitted, 1.5.2.11.1 Results of examination of individual seeds
see 6.5. or seedlings
– If individual seeds are tested at the end of the germi-
nation test, the result must be reported in accordance Suggested phrases for reporting divergent seeds or seed-
with 5.9. lings depending upon the result are as follows:
a. if none was found: ‘The test performed revealed noth-
In addition, in the case of species of Fabaceae, one of the ing to indicate that the species (and/or variety) stated
following, and only one, must be reported: by the applicant is incorrect.’
either (in cases where the percentage of the viability of b. if non-conforming seeds were found: ‘Out of … seeds
hard seed is not determined) ‘Tetrazolium test: ...% of examined, … seeds do not conform to the authentic
seeds were viable, ...% of hard seeds found in the test’ standard sample of the species (and/or variety) stated
or (in cases where the percentage of the viability of hard by the applicant.’
seed is determined) ‘Tetrazolium test: ...% of seeds c. if non-conforming seedlings were found: ‘Out of ....
were viable, ...% of hard seeds included in the percent- seeds producing normal seedlings, …% do not con-
age of viable seed’ form to the authentic standard sample of the species
(and/or variety) stated by the applicant.’
d. if the total working sample was found to be of a spe-
1.5.2.10 Seed health test cies and/or variety other than that stated by the appli-
cant: ‘The sample does not conform to the authentic
The results of a test for seed health must be reported under standard sample of the species (and/or variety) stated
‘Other determinations’ as follows: by the applicant.’
– either qualitative or quantitative results, as specified in
the individual methods;
– negative and positive results, as specified in the indi-
Chapter 1: ISTA Certificates
vidual methods;
– the scientific name of the pathogen detected;
– the percentage of infected seeds;
– the method used, including any pretreatment (7.2.2);
– the size of the sample or fraction examined;
– any additional permitted procedure used.
The result of a field plot examination must, whenever The result of an excised embryo test must be reported un-
possible, be reported as a percentage of each other spe- der ‘Other determinations’ as follows: ‘Excised embryo
cies, other variety or aberrant found. When the expression test: ……% of seeds had viable embryos’
of the result as a percentage is not possible, appropriate Further details may be given at the discretion of the
comments regarding the conformity of the sample may be seed testing laboratory, e.g. percentages of seeds that were
reported. empty, insect-damaged or physically damaged.
If nothing worthy of special comment was found the
following statement is suggested: ‘The results of a field
plot examination of this sample revealed nothing to indi- 1.5.2.15 Weighed replicates
cate that the species (and/or variety) stated by the sender
is (are) incorrect.’ The result of a weighed replicates test must be reported in
the space provided as follows:
– The result of the purity test (if requested), in the spaces
1.5.2.12 Moisture content provided for purity tests.
– ‘N’ must be entered in all the spaces provided for re-
This Rule is applicable to both the oven method (9.2.7) porting the percentages of the components of the ger-
and the moisture meter method (9.3.2.7). mination tests.
The result of a moisture content test must be reported
in the space provided to the nearest 0.1 %. The following additional information must also be report-
ed under ‘Other determinations’:
The following additional information must also be report- – average weight of four replicates;
ed under ‘Other Determinations’: – average number of normal seedlings in four
– For the oven method (9.2.7), the method (i.e. duration replicates;
and temperature) must be reported. – number of normal seedlings per kilogram;
– For the moisture meter method (9.3.2.7), the state- – other information as specified in 1.5.2.6 and 5.9.
ment: ‘A moisture meter was used’ must be entered.
– If germinating seeds were present in the sample, the Upon request, other seeds found to be present in the
following statement must be entered: ‘Germinating weighed replicates may be reported, giving the scientific
seeds were found in the submitted moisture sample.’ name(s) and number(s) of seeds found.
– If mouldy seeds were present in the sample, the fol-
lowing statement must be entered: ‘Mouldy seeds
were found in the submitted moisture sample.’ 1.5.2.16 X-ray test
– In the case of pelleted seeds (see Chapter 11), the fol-
lowing statement must be entered: ‘The seeds of the The results of an X-ray test must be reported under ‘Other
submitted moisture sample were pelleted, and the determinations’ as percentages of filled, empty, insect-
moisture content reported is the average of seed and damaged or physically damaged seeds, as follows:
pelleting materials.’ X-ray test results
– For Arachis hypogaea, one of the following statements ……% filled
must be entered: ‘The submitted sample for moisture ……% empty
determination consisted of seeds in their pod’ or ‘The ……% insect-damaged
submitted sample for moisture determination consist- ……% physically damaged
Chapter 1: ISTA Certificates
– The seed moisture content before the test must be re- b. Conductivity test after deterioration
ported. Where the moisture content has been adjusted – The result must be expressed in μS cm–1 g–1 to the near-
before the test, both the initial moisture content and the est 0.1 μS cm–1 g–1.
calculated moisture content after adjustment must be – The results must be accompanied by a statement of the
reported. specific variables used during deterioration (method
– The results must be accompanied by a statement of the used to raise seed moisture content, raised seed mois-
specific variables used in the test (soaking time and ture content, deterioration period and temperature) and
temperature) in the conductivity test (soaking time and temperature).
The result of a seed vigour test using the accelerated aging The result of a seed vigour test using the radicle emer-
(AA) method must be reported under ‘Other determina- gence test must be reported under ‘Other Determinations’
tions’ as follows: as follows:
– Results are expressed as a percentage, calculated to – Results are reported as a percentage of seeds with
the nearest whole number (5.8.1) of normal seedlings, emerged radicles calculated to the nearest whole num-
abnormal seedlings, hard seeds, fresh seeds and dead ber (5.8.1). If the result is found to be nil, it must be
seeds. If the result for any of these categories is found entered as ‘0’.
to be zero, it must be reported as ‘0’. – The results must be accompanied by a statement of the
– The seed moisture content before the test must be re- temperature used for the test and the time of the radicle
ported. Where the moisture content has been adjusted emergence counts in hours, e.g. ‘Radicle emergence
before the test, both the initial moisture content and the test 90 % with emerged radicles after 66 h at 20 °C.’
calculated moisture content after adjustment must be
reported.
– The results must be accompanied by a statement of 1.5.2.17.5 Tetrazolium vigour test
the specific variables used in the test (seed weight per
AA box both before and after ageing, ageing time and The result of a seed vigour test using the TZ method must
temperature). be reported under ‘Other determinations’. Results are ex-
pressed as a percentage, calculated to the nearest whole
number of vigorous seeds, e.g.: ‘Tetrazolium vigour tests
1.5.2.17.3 Controlled deterioration test using 0.1 % TZ solution for 3 h at 35 °C: 90 % vigorous
seeds’.
The result of a seed vigour test using the controlled dete-
rioration test method must be reported under ‘Other deter-
minations’ as follows for the two alternative methods of 1.5.2.18 Size and grading of seeds
assessing deterioration in 15.8.3.4.3:
The result of a screening analysis test for size and grading
a. CD germination test of seeds must be reported under ‘Other determinations’ as
– Results are expressed as a percentage, calculated to the the average of two screening analyses falling within the
nearest whole number (5.8.1), and stated as ‘Total ger- permitted tolerance limits.
minated seeds (normal plus abnormal seedlings) … %’
and ‘Normal seedlings … %’. If the result for either of
Chapter 1: ISTA Certificates
1.5.2.19 Weighted average test for seed lots 2. The percentage by weight of mixture components,
transported loose in bulk containers pure seeds or inert material according to declaration
using the sum of the weights of the pure seed fraction
The result of a weighted average test performed on seed and the declared inert material.
lots, as described in Chapter 17, must be reported in the 3. The percentage by number of the pure seeds of the
normal way, except that: mixture components using the total number of seeds
a. Across the date of sampling, date sample received, of the pure seed fraction.
date test concluded and test number boxes insert the
statement: ‘Seed loose in bulk container(s) – see under In addition, if applicable, the percentage by weight of the
Other determinations.’ ‘inert material according to declaration’ using the sum of
b. Under ‘Other determinations’, list the test number, the weights of all mixture components must be given to
date of sampling and date test concluded of all constit- one decimal place under ‘Other determinations’.
uent lots together with the statement: ‘The test results
reported represent the weighted average of the results
reported on these certificates which were not signifi- 1.5.2.20.2 Determination of other seeds by number
cantly different from each other.’
The results of a determination of other seeds by number
on a seed mixture must be reported according to 4.7.
1.5.2.20 Seed mixtures
The results of tests on seed mixtures can only be report- 1.5.2.20.3 Germination, seed viability, seed vigour and
ed on a Blue International Seed Sample Certificate (see other tests using replicates of 100 seeds
1.2.2).
For the species tested, ‘Seed mixture’ together with the Test results are reported only for those species for which
mixture composition according to the declaration of the methods are given in the appropriate Chapter of the ISTA
applicant, must be entered. Rules. The results of these tests must be reported under
‘Other determinations’.
Germination test results are not reported in the ‘Ger-
1.5.2.20.1 Purity and component analysis mination’ section of the certificate (an ‘N’ must be entered
there), but under ‘Other determinations’. When 100 or
The results of the purity analysis are reported according more seeds are tested, the percentage results of the test for
to Chapter 3. each mixture component tested are reported to the nearest
The actual weight of sample examined to the minimum whole number. The number of seeds tested is also report-
number of decimal places indicated in Table 4.1 must be ed. Tolerances as described in the appropriate Chapters
reported under ‘Other determinations’, i.e. ‘Purity and are applied to tests of 400, 300, 200 and 100 seeds.
composition analysis: … g of seed examined.’ When fewer than 100 seeds are tested, the actual num-
ber of seeds in each category (e.g. normal seedlings or
The mixture composition is reported under ‘Other deter- viable seeds) is reported, together with the total number
minations’ in one of the following formats, as requested of seeds tested.
by the applicant: The method used in the test must be reported on the
1. The percentage by weight of the pure seeds of the certificate according to the appropriate Chapter for each
mixture components using the total weight of the pure component species tested.
seed fraction. In addition, if applicable, the percentage
Chapter 1: ISTA Certificates